Peptide‐Templated Acyl Transfer: A Chemical Method for the Labeling of Membrane Proteins on Live Cells1. Issue 38 (31st July 2014)

• Record Type: Journal Article
• Authors: Reinhardt, Ulrike; Lotze, Jonathan; Zernia, Sarah; Mörl, Karin; Beck‐Sickinger, Annette G.; Seitz, Oliver
• Is Part Of: Angewandte Chemie international edition. Volume 53:Issue 38(2014)
• Page Start: 10237
• Page End: 10241
• Publication Date: 2014-07-31
• Subjects: Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201403214
• Languages: English
• ISSNs: 1433-7851

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>The development of a method is described for the chemical labeling of proteins which occurs with high target specificity, proceeds within seconds to minutes, and offers a free choice of the reporter group. The method relies upon the use of peptide templates, which align a thioester and an N‐terminal cysteinyl residue such that an acyl transfer reaction is facilitated at nanomolar concentrations. The protein of interest is N‐terminally tagged with a 22 aa long Cys‐E3 peptide (acceptor), which is capable of forming a coiled‐coil with a reporter‐armed K3 peptide (donor). This triggers the transfer of the reporter to the acceptor on the target protein. Because ligation of the two interacting peptides is avoided, the mass increase at the protein of interest is minimal. The method is exemplified by the rapid fluorescent labeling and fluorescence microscopic imaging of the human Y₂ receptor on living cells.</p> </abstract>